speaker: Greg Fahy

talk title: Thymus rejuvenation progress update

date: 2021-05-27

video: https://www.youtube.com/watch?v=KukRAXnNOJM&list=PLH78wfbGI1x0Bpmq40HEVtYGj_CsEfHSk&index=117

https://interveneimmune.com/

LLM summary: Greg Fahy presented a comprehensive update on the TRIIM (?Thymus Regeneration, Immunorestoration, and Insulin Mitigation) trial and its extension, TRIIM-X, demonstrating thymus regeneration in middle-aged and elderly humans via intermittent administration of human growth hormone (hGH), DHEA, and metformin. TRIIM achieved significant thymic fat reduction, naive T-cell increases, epigenetic age reversal across multiple clocks (e.g., 2.5 years via PhenoAge, Hannum, GrimAge), and ancillary benefits like improved kidney function and reduced inflammation, without elevating cancer risk. TRIIM-X replicates these in a larger, mixed-gender cohort (40-80 years, n≈23 ongoing, targeting ~80) at the Lundquist Institute, with preliminary data showing anti-inflammatory effects, PSA reductions, lipid improvements, eGFR gains, pulmonary enhancements, and rapid PhenoAge plasma clock reversals (e.g., 4 years in months), alongside persistence of TRIIM benefits six years post-treatment. Mechanistic plausibility stems from thymic grafts reversing non-immune aging hallmarks (e.g., hepatic tetraploidy, insulin sensitivity), monocyte depletion restoring NAD+, and complementary anabolic actions countering catabolism, with future directions including proprietary hGH formulations, thymic reprogramming for transplant tolerance/autoimmunity, and scaled self-funding trials.

Opening and Introduction

So Greg, we're incredibly excited. Thank you so, so much for joining. I'm super excited for a progress update on what you've been up to in terms of thymus regeneration.

Okay, well thank you. Thank you very much, Alison. It's great to see everybody. I see that among our audience members, we have at least one volunteer in the TRIIM-X trial. So if I lie to you, she will straighten me out on this.

So I'm going to share my screen if we can manage to do that. I actually have a presentation to give to you this time and let's see if we can manage to make this work.

Okay so Alison said that this doesn't really need an introduction but I'm going to give you an introduction anyway just to make sure everybody's sort of up to speed on this. We start off from the same basic point and by the way thank you Alison for your invitation. This is a special group and it's always a pleasure to interact with this group.

Importance of the Thymus in Aging

Okay, so you know all of this, but this is different graphics than I usually show, so I'm going to show this to you anyway just to make sure that there's no stragglers in the audience. It's not quite up to speed on this.

So the general idea that we started off with is that the thymus is very important and it withers away with age. And so you can see in the diagram on the bottom, the pink area would be the functional thymic mass and the white area would be fat. And by the time you get up to 70 years of age, there's very little left in the way of functional thymic mass. And that is diagrammed in terms of how it affects the anatomy on the bottom of that figure.

The reason that this is important is that we die because of this. And I've shown figures before about how mortality rate goes up as immune system function goes down. I'm not going to even repeat those for you. And you all know about COVID. I don't have to go through that again with you.

So the idea is that we have to do something about this so that we can prevent our T cell population from being depleted and loused up by the slings and arrows of outrageous aging.

So that's where we started. And as you all know the thymus is in your chest cavity. And the beauty of this approach to aging is that it is doable.

Reversing Thymic Involution

So thymic involution which is what this process of deterioration of the thymus is can be reversed in many different ways. And there's a list of, you know, some of the ways that exist out there on the left. But I've highlighted the first item here, growth hormone or somatropin because it's my favorite way of regenerating the thymus. And it's my favorite way for all kinds of reasons.

One of those reasons is that it has been proven effective in many different species. So in rats, Keith Kelly in 1986 showed that you can regenerate the thymus both structurally and functionally in rats. And later on, it was shown that you can regenerate the thymus of the dog using growth hormone. And there's even a cat study showing that giving IGF-1 to cats, which is what is increased by growth hormone, radically changes the structure of the cat thymus. The cortex, it's all pale because depleted of cells on the right and the old cat thymus and the rejuvenated cat thymus, everything comes back to normal.

And then before we came along was even a few studies on HIV victims using CT to show an improvement in thymic structure.

So that's one reason.

Do dogs have their own special growth hormone that's a different sequence? I believe that they do. I haven't investigated that specifically, but presumably they do. And that's actually a key question because I'm also working with different people on a dog aging intervention study. And one reason that I'm not really thinking about trying TRIIM on dogs right now is that I don't know how to get dog growth hormone in any kind of serious way. But if anybody has an idea about that, that would be great.

Having said that, there are a lot of studies in which human growth hormone has been given to animals and been shown to be biologically active, but I'm afraid that that may lead to some side effects. And so I think it's probably not the way to go.

So this is one reason that I like growth hormone is that it seems to work across the board.

Additional Reasons for Preferring Growth Hormone

But there are other reasons too which some of you are familiar with. One of them of course is that has to do with safety. You know there's a long clinical experience giving growth hormone to growth hormone deficient adults since 1996. It's pretty safe. The side effects are known and they're reversible and mild.

And it has in addition to not having too many bad side effects a lot of positive effects including accelerated wound healing, helping with learning and memory, you know, promoting various good processes in your brain, like making new synapses, new neurons, opposing Alzheimer's disease, and then in the periphery, growing cartilage, which might help to forestall arthritis as we get older.

And what we're illustrating in this figure is, it's the last couple points, this profound effect of building up lean body mass and burning fat. So you can see what happens to us is control. So if we're not treated with growth hormone, lean body mass goes down and the mass of your liver and your spleen and your kidney and your muscle, it all goes down with age. The only thing that goes up with is fat, which is just the thing that most of us are not that fond of having increased with age.

But if you start growth hormone and keep it going for a year, all of these trends tend to reverse. This is really a deep, deep biological, profound change. And as part of this, if you're growing back your liver and you're growing back your muscle and you're growing back your skin, it turns out you're also growing back your thymus. And that's really the thing that people have overlooked for the most part in the past is that key part, the thymus, which you need in order to stay alive.

Managing Growth Hormone Side Effects

So there is a particular side effect that I was concerned about, which is that growth hormone induces the production of insulin. And that is a pro-aging factor. But I was able to show many years ago that that can be blocked by co-administering growth hormone in DHEA. And the good thing about that is that DHEA in and of itself has many different anti-aging features. It reduces all cause mortality et cetera et cetera. It also known as the neurosteroid. It's present in the brain. It probably helps protect against brain aging like growth hormone does. And some people when they taking DHEA especially women, tend to experience an improvement in mood and that sort of thing, perhaps related to its actions in the brain.

But sometimes DHEA is not enough to counteract the diabetogenic effect of growth hormone, this increase in insulin. And for that, we add in metformin, which has another number of advantages. You guys have heard Nir Barzilai speak about the virtues of metformin. It's an insulin sensitizer, so it strikes at the heart of this problem. It also has a lot of other beneficial effects.

So I think this is a winning combination. And all of these interventions are obviously pretty powerful. In combination, the sum of the combination is more than the sum of its parts, basically, which helps to explain why we got some of the good results we got, I think.

Greg, very quick question. Am I missing a question? Yeah, very quick one. Sure. Do you know that if one monitors with a continuous glucose monitor, which is easier than monitoring insulin, how does that affect this? Like, do you think that metformin and DHEA are just so good that even if you don't have an insulin problem, it's a win? Or do you think that this is really mostly because of the anti-insulinogenic features of those treatments?

Yeah, I know that DHEA has effects that are in addition to its anti-insulin effects. Metformin may have as well. But obviously, metformin was developed to be an anti-diabetic drug. And that's probably one of its major foci of action. But I think that DHEA, its effect on insulin is only present if you're raising insulin with growth hormone. It seems to be particularly aimed at that particular endpoint.

There's one study that indicates that DHEA can lower insulin in ordinary people, but the vast majority of studies say that it cannot. So the only case in which I think is really reliable in reducing insulin is when insulin is raised by growth hormone. I think that that's not a coincidence. That's the reason that I proposed DHEA in the first place is that when we're young, we have plenty of growth hormone, but we're not diabetics. And so I think the reason for that is that we also have plenty of DHEA. So by combining the two, DHEA naturally and physiologically opposes this diabetogenic effect of growth hormone, whereas metformin is just there as a general purpose insulin sensitizer that helps without having any built-in physiology behind it.

Okay, let me ask it a slightly different way very quickly. Okay. Does growth hormone increase insulin without increasing blood sugar? Well, it depends. It's a matter of degree. So the first thing that happens is the insulin goes up. And if you keep increasing the dose of growth hormone too much, eventually the increase in insulin is not enough to overcome the insulin resistance that's induced by the growth hormone. And then at that point, your glucose can go up as well. So both of them will go up, but it's a matter of dose. Thank you. Okay, you're welcome.

Overview of the TRIIM Trial

All right. So that's kind of the background. And because of all that background, as we all know, we did the TRIIM trial based at Stanford University. It was held from 2014 to 2015. There were two cohorts. The first cohort was the larger one, which you see in the center. Second cohort was just three guys, and you see them on the right there.

And the aim was to regenerate the thymus using this novel combination of drugs called growth hormone, DHEA, and metformin. The FDA considered that a new drug entity so we had to get an IND from the FDA to do this study but we did that. We only had nine in the end. We actually had a 10th guy who dropped out after about a month due to anxiety primarily. He never actually should have been in the trial. He had a high familial cancer risk, and he was worried about taking growth hormone, but he wanted to get his thymus regenerated. So he entered the trial in the first place, but then thought better of it after a month and dropped out.

So the treatment is you give this combination, this cocktail, four times a week for a year. And the items are juggled and optimized for each individual frequently throughout the course of that one year. And as you know, we published this in September of 2019, and it got a lot of attention.

So the trial was pretty successful. Even though it was only intended to be a pilot study and a dose ranging study, it turns out all kinds of interesting things came out of this trial.

Key TRIIM Results: Safety, Immunology, and Unexpected Benefits

And here are a few of them related to safety and immunology. The first thing was that we did regenerate the thymus, as you know. And we documented that by disappearance of thymic fat. But since the thymus is located under the breastbone, the sternum also has fat in the bone marrow of the sternum. So we looked at the regression of fat in the sternum as well, and we saw regression of fat in the bone marrow of the sternum as well. But it wasn't nearly as profound as what we saw in the thymus, probably because we're not just burning fat, we're actually regenerating functional thymic tissue.

And evidence for that is shown in the middle set of figures on the left, showing an increase in naive T cells and recent thymic emigrants. And then below that, in terms of safety, we saw several indications that cancer risk was actually going down, even though when you're given growth factors like growth hormone, you're always worried about whether you might be promoting cancer.

But we saw a reduction in PD-1 cells, which are cells that are basically incompetent and they prevent your body from attacking cancer. And those declined with time. We also saw a decline in prostate antigen which is a marker of cancer risk in the prostate. There's a spike in the middle there because some of our guys got a little out of control and engaged in sexual activity before they got their blood drawn about six months into the trial and so that screwed up the assay. But as soon as we corrected that behavior by telling them not to do that right before getting their blood drawn, everything went right back down to normal. So that's kind of another aspect of the treatment, which is a good side effect, actually.

And then there's something called the lymphocyte to monocyte ratio, which correlates with all kinds of cancers. And a high ratio is good, and our ratios went up, at least based on the way that this ratio was measured at Stanford University with their sophisticated CyTOF measurement techniques.

So we felt very reassured that cancer risk was not being exacerbated.

And then in terms of other immune aspects, C-reactive protein actually decreased in the trial, which is a sign that we're not increasing inflammation. Globally speaking, we're actually decreasing inflammation, at least as sensed by CRP.

And then the last thing was insulin. So we gave metformin and DHEA to control insulin levels. And we were not completely successful at that. But we did suppress insulin fairly well. And most people in the trial had insulin levels below the median for the general population, even in the face of all this growth hormone that we're giving them, except for one guy whose insulin went up at the very end of the trial and kind of screwed up our statistics. But generally speaking, it was satisfactory.

And then unexpected results that you've probably heard of. There was one guy in the trial in particular who had his hair start getting darker. In the course of the trial, his wife pointed it out to him. So we started paying attention to him. And we have several photographs of this change. A couple of other people in trial seem to have had the same effect, but we didn't have the chance to document them quite as well.

A couple of things that we were not expecting is that the estimated glomerular filtration rate the filtration rate of the kidneys actually went up over time. And that was statistically significant in different time points and overall. And seemed like it might have even been continuing after the treatment ended.

And then the last thing here is that this increase in lymphocyte to monocyte ratio, which I mentioned to you before, is really driven mostly by a reduction in monocytes. And monocytes actually destroy tissue NAD. And depleting tissue NAD is a great way to get yourself aged. So reducing these cells might be one reason that we saw some of these benefits would seem to go beyond just immune system aging and makes you wonder if there was a global aging reversal.

And as you all know now...

Epigenetic Age Reversal in TRIIM

So that's what we saw. So just to summarize this for you, we measured the epigenetic age based on four different epigenetic aging clocks before treatment and during the course of treatment. And we measured the change that took place as the difference in epigenetic age minus age during the treatment compared to the epigenetic age minus age before treatment. So the epigenetic age minus age before treatment will vary from every individual. But if we subtract that for each individual from what's going on in time, we isolate the effect of treatment independent of somebody's background.

As you can see in every case, based on DNA methylation age, the PhenoAge, Hannum's aging clock, and the GrimAge clock, epigenetic age went in reverse. And you could detect it at nine months into the trial. And you could also detect it at 12 months, even more so. And what you see in panel E is an average of all of these results, which are measuring epigenetic age in different ways.

And then, as you know, probably, if you look at the difference in slopes from zero to nine months versus nine to 12 months and do a statistical test on the slope, you find that it's statistically significant, which means that the epigenetic aging reversal effect was accelerating over time. And you're going about four times faster in the last three months of the trial than you were in the first nine months.

So it takes the body a while to respond and realize it's supposed to be younger than it is.

Is Epigenetic Reversal Real? Mechanistic Plausibility

So the question that we're left with as a result of all of this, is this real? Is this some kind of a fluke? Is this some kind of error on our part? Or is this actually a real phenomenon. So the rest of the talk will be pretty much devoted to looking at that question from several different angles.

So you may have seen this figure before. I've shown this several times. I'm very impressed by this data. This is a compilation of results from five different individual experiments that were done and published separately, and this all sort of put together.

So it's been shown in the past that if you give a thymus transplant to an old animal, you can improve the crosstalk between brain cells through beta receptors found in the brain, because the beta receptor population plummets with aging and it's restored by a thymus transplant. And you can lower plasma insulin levels. So it seems like the thymus somehow helps insulin sensitivity as well.

And in the liver, as you get old and you happen to be a rat or a mouse, you develop this condition of tetraploidy. A lot of liver cells have twice as much DNA as they're supposed to, and that's actually reversed by a thymus transplant.

So we knew that thymus regeneration seems to have the ability to reverse non-immunological aspects of aging, and that's kind of what we saw in TRIIM. So that's consistent with our study actually having a real effect.

But it's not just the thymus because if we actually correlate epigenetic age reversal with a reversal of thymic involution, we find that there's no statistical significant correlation. So we think that there's two independent effects that may be driving aging reversal in our volunteers.

We mentioned that lowering the monocyte level may restore tissue NAD and if you restore tissue NAD in mice you can reverse mitochondrial aging and other aspect of aging and postpone the onset of age mortality. So that's another possible reason why our effects can be sort of considered to be plausible.

And then, as I mentioned before, each one of these agents that we're using has been shown to oppose aging in various ways, probably to a large extent in complementary ways. So combining them hits many different anti-aging pathways. And this catabolic effect of aging is being opposed by the anabolic effects of both growth hormone and DHEA.

So it's kind of believable that it might have some profound effect on aging.

And I just point out in passing here that we've all heard about heterochronic parabiosis, and there's debate about whether it's youth factors going from the young to the old animal or old age factors just being diluted from the old animal. Probably they're both true. But last time I spoke to Irina Conboy about this, I kind of convinced her that the evidence is supportive of the youth factor idea, you know, despite her arguments to the contrary. And I just have to point out that in youth, you have lots of growth hormone and DHEA. So that might be part of the effect of this heterochronic parabiosis effect, benefiting the older parabiont.

New Evidence from PhenoAge Plasma Clock

But here's some new evidence that you haven't seen before. So Steve Horvath decided to go back and retrospectively look at the TRIIM data using a different measure of aging. So we looked at the PhenoAge clock, epigenetic aging clock before. But that requires that you take DNA samples and that you measure methylation sites and that you wait until the end of the trial. And so you don't get results during the trial itself.

But there is a component to the PhenoAge clock, which relies only on testing things that are available in plasma or blood. Some of the white cell populations are red cell populations in the blood. And so Steve went back and recalculated this plasma aspect of PhenoAge for guys in the TRIIM trial and found that on average just as we found with the epigenetic clocks time going back by two and a half years in the PhenoAge plasma clock time went back by two and a half years as well.

And these are just our best four examples, probably from that retrospective analysis, showing PhenoAge going backwards in time using this fifth way of estimating biological age.

So that kind of gives us even more confidence of what we saw based on the original four aging clocks. We now have a fifth aging clock that we can add to that. And it has the advantage of being able to be something we can check on in real time. So we don't have to wait until the end of the trial to get some idea what may be going on. And we're actually doing that in TRIIM-X, as you'll see in a bit.

And I just will say parenthetically on the left, what you see there is the PhenoAge versus the actual age, the chronological age. And all the white dots are people who were in the TRIIM trial at different stages of being in the trial. And the line, the identity line is that heavy black line. And you can see everybody in the trial was pretty much younger biologically than they were chronologically at the start of the trial. And yet we can see this reversal of aging based on the PhenoAge clock in spite of that, which also agrees with what we saw with the epigenetic aging clocks.

Introduction to TRIIM-X Trial

Okay. So I've given you a lot of hand-waving about, gee, you know, We think that this is real. We think that all kinds of reason to believe our results. But the only way to really know is to repeat the experiment. So that's what we're talking about for the rest of this presentation. And it's the main point of this talk, since it's the part you haven't heard about before too much.

So we have launched something called the TRIIM-X trial, X standing for extension of the original TRIIM trial. We're repeating it and we're extending it in a number of ways. It's on clinicaltrials.gov if you want to read about it. And it includes both men and women now with an expanded age range from 40 to 80 years of age. So far we have not enrolled anybody less than 50 but we have enrolled people right up to the age of 80. In fact we got one guy in who formally qualified for the trial about one day before he turned 81. So he just managed to squeak in by the skin of his teeth.

But we have been starting this and the trial is now headquartered local to us for convenience. It's at the Lundquist Institute. I'll tell you a little bit more about Lundquist in a second. And you can see our first cohort of three people at the end of November of last year, standing there in front of our headquarters building at the Lundquist Institute with Bobby Brooke, our CEO, next to them.

TRIIM-X Infrastructure and Enrollment

Okay, so I used to know about something called Harbor UCLA. It was it's a big medical complex in Torrance, California, but it's part of this larger medical and research complex of buildings and structures. And somewhere within that big general medical research complex, there's something called the Lundquist Institute, which has several component buildings to it, which are sort of blown up and magnified on the right.

So the upper right of that figure, there's a building called the Medical Research Laboratory, also called the Biolabs. This is where our corporate headquarters is located, and it's where we give seminars to cohorts that come in for the trial.

We have two modes of signing up for the trial. You can either come in person if you want to have some specialty tests done, or you can participate entirely by telemedicine if you wish, if you don't want to have these extra tests done. But we give informed consent. We give seminars. We can have medical exams done either in person or by telemedicine at the bio labs.

Now, nearby the bio labs are three other buildings of interest. The first one, which is highlighted in the green box on the left side, is a building where we can get DEXA scanning for body composition, your lean body mass before and after treatment. And also this is where you go to get your blood drawn. By the way, everybody participating in the trial has had to get a COVID-19 test as well to make sure that you're not infectious before being on the premises of the campus.

We also, in addition to offering people these body composition scanning, we offer them CT imaging of the thymus at heart so we can look at thymus regeneration. And we have had a number of people take us up on this, including some of the women in the trial, because we have no data on thymus regeneration in women so far. Actually, nobody does. So we're going to get the first evidence ever on that score.

And we can also image coronary artery calcification, which correlates with the risk of death and disability. And we don't know if that will be affected by our treatment, but it would be fantastic if it were affected by our treatment.

And then the last building is where you can get cardiopulmonary exercise testing done and leg strength examination. We want to know if you get stronger as a result of being in the treatment and whether your exercise capacity improves because that's a measure of frailty and frailty, of course, is not good for anything.

Okay. So this kind of shows the course of the enrollment activity so far. We began on Thanksgiving week in November of last year and the total number of enrollees has been going up pretty steadily ever since. We only have five women in the trial at the moment.

You notice that there's a dip in the enrollment numbers around month three. That's because we had two people in the trial who were sex hormone enthusiasts and decided after passing their enrollment blood exam that they wanted to tank their testosterone and estrogen levels up to above the normal range, like vastly above the normal range. And we pointed out to them that that will involute their thymuses and destroy the effect of the treatment, but they didn't want to hear that. So they eventually dropped out after about a month or so. But everybody else has stayed in the trial.

And as you see by the black lines at the end of these curves we expecting right before the end of the sixth month which is May that we have another cohort of one added onto the list. So we're still continuing to enroll people. Our target for enrollment is on the order of 80 people something like that. In TRIIM we had nine people as you know here we're up to about 23 and so it's been an interesting challenge for me because this is a very customized hands-on kind of treatment protocol and so we've had to work out ways to handle that.

TRIIM-X Protocol Management

So this schedule for the week has been uniformatized for almost everybody. And of course, there are exceptions sometimes as we go along, but we start here with Monday. So we usually, if we want to test how people in the trial are doing, we have their blood drawn on Monday. They don't get an injection that day, but they get injected on Tuesday night. And by drawing on Monday, we have plenty of time for the results to come in soon enough for me to look them over and contact the volunteer on Saturday and Sunday and let them know how we want to change their doses for the next time interval of the trial.

So usually on Sunday is when the doses are updated and new doses are tried out, basically. Um but we we had to do it this way because if we uh draw blood on thursday or friday uh sometimes they do not get uh analyzed quickly because of the weekend so we we measure some esoteric things that have to be shipped between different laboratories and sometimes that creates problems so by doing it this way uh we can update everybody on the same schedule so i We only have to work on updating people basically on Friday, Saturday, and Sunday, and mostly on Saturday and Sunday. So that usually burns my entire weekend, every single week.

So we see how much we can expand this population. But thanks to Bobby's brilliance computing we able to download the blood chemistry results into a specialized Excel file that takes all of the data puts it where we want it, and then allows me to copy it into specialty files for each individual so I can track how each individual is doing with respect to all kinds of different parameters and ratios of parameters and and all kinds of things so that I don't have to manually transcribe the data like I had to before, which is a huge help.

And we're still working on algorithms to help me with the dose updating process, but it turns out to be fiendishly complicated and it's not going to be all that easy to reduce this to an algorithm, but we're starting to do that now.

So that's how we're managing the practicalities of the trial.

Volunteer Experiences in TRIIM-X

And just to tell you how things are going in general, how the volunteers are reacting to the trial. Some people have said that they can feel themselves getting younger. Some people have said that their massage therapists tell them that they are more ripped than they've ever been. People feel more energetic. I've actually experienced that myself because I'm one of the guinea pigs in this trial myself. I finally enrolled myself in the trial, and so I can attest to this sort of thing.

Some people have reported joint pains, but so far nothing has gotten to the point of making anybody have to back off on their doses or drop out of the trial. Even though joint pains are a known side effect of growth hormone administration, it's because it wants to grow you. And that includes growing your bones. But of course, after puberty, you cannot grow your bones. So in the short run, you may have some arthritis type pain. But my theory is that in the long run, you're actually better off because you're restoring cartilage that we tend to lose with age. And if you have more cartilage, then in the long that should be beneficial for you.

So nobody's dropped out due to side effects. We did have those two dropouts for the reasons that I described.

I have to tell you also that sort of similar to the experience that we had at TRIIM, the TRIIM-X volunteers are extremely smart. They very health conscious. They flood me with information mostly about themselves. A lot of them are biohackers and they have been tracking themselves for years and years and they have reams of data. And some of them send us data that they're collecting parallel to the trial through other doctors and other lab, clinical chemistry labs.

So we got plenty of data. In fact, some of the volunteers actually asking us to run more assays on them, which is pretty cool because it allows us to look at things that we otherwise wouldn't be able to look at. And that's actually been illuminating in many different ways. I'm learning all kinds of new things about female sex hormones that I didn't know before and how they're processed and all kinds of stuff.

So it's a learning experience. We also have a lot of people in this trial that are not in such great health. We have people with bowel conditions. We have people with borderline kidney failure, you know, the not in kidney failure, because we don't allow that, but people are kind of on the edge. And so, and we have, you know, pretty old people and pretty skinny people and all kinds of people in here that we didn't have in TRIIM.

So we're learning all kinds of new stuff about how to handle these cases and how these cases respond. So it's really a positive experience.

Preliminary TRIIM-X Results: Replications

And then I'm going to spend the rest of this presentation talking about preliminary results. Now, as I've shown you, we're only less than six months into this trial so far, and that's just for the first three people in the trial. Most people are in it for less than that, and we have a year to go to finish the first cohort, and we'll have a year to go after we enroll the last cohort, so our final results are going to take years to come in, but we do have preliminary results, and some of them are interesting enough to share with you.

And some of them are expected, meaning that they're actually replicating what we saw in TRIIM, which is the whole idea of TRIIM-X is to see if we can repeat some of the things that we saw before, but we've also seen some really interesting things, in my view, that are unexpected and new, and so I'm going to share those results with you.

So one of the things that we were hoping to replicate is this effect on CRP, C-reactive protein. People have worried that regenerating the thymus and restoring immune system function might be bad because one of the things that happens with aging is you have inflammation. And so if you augment immune functions, you might increase inflammation. But in TRIIM, we did not see that. We saw a decrease in C-reactive protein.

And I'm just going to be showing in these figures, I'm taking three exemplars and plotting their results out. So the first three time points, generally speaking, are baseline data. So we have our pre-screening data point. And then when they enroll during the enrollment week, we take two more baseline samples. So generally speaking, those three samples are in agreement with each other. So that forms sort of the baseline for everybody.

And then once you get beyond the vertical dotted line, now you're into treatment time. And I'm not specifying exactly what the time points are here, just to keep things simple. But what you can see in this case is that for two males and one female in these particular examples, you can see that C-reactive protein is holding out fairly constant between one and two before treatment and then falling in each case after treatment.

So that seems like preliminary evidence that we're actually reproducing this anti-inflammatory effect of the treatment.

Another preliminary result that is consistent with the TRIIM result is improvements in kidney function. It's one of the more extraordinary results of TRIIM is that normally as we get older, our kidneys always get worse. But in TRIIM, the kidneys seem to get better. And we're seeing that in TRIIM-X as well. In this case, these are all males, but you can see that the trend is pretty distinctly upward and pretty quickly for the most part in all three of these cases.

Uh we also saw in TRIIM an apparent improvement in prostate health meaning a reduction in PSA level and an increase in the percent free PSA we're seeing both effects in TRIIM-X so that seems to be reproducing the TRIIM results as well um and these are just three examples uh uh one person who had a relatively high PSA actually above the normal range but had prostatitis so we knew it wasn cancer starting to reverse pretty significantly after a couple treatments or a couple of intervals of treatment.

And the other people starting off with pretty low PSAs, but still improving, which we also saw in TRIIM. We saw in TRIIM that regardless of what your baseline PSA was, it always went down no matter what. So these are pretty encouraging signs that we're able to reproduce at least some of the aspects of the TRIIM trial.

Preliminary TRIIM-X Results: New Observations

Here's the beginning of some new things that we've seen. So these two people showed a reduction in their triglycerides. Normally, I think what the trend is going to turn out to be, although we don't have all the data yet, so we can't speak to this definitively yet. But I think that what we're generally going to see for most people is that as you begin treatment and you begin breaking down fat from your fat stores, your triglyceride, your blood triglyceride levels tend to come up. And then as you sort of take that to a point of completion or steady state, then it will come back down again.

But in these two cases, the triglyceride levels came down right away. And in one case, pretty profound. So you'd like your triglyceride levels to be lower rather than higher. And so this is an interesting new observation that may pertain to a subpopulation of people in the trial.

Similarly, we like our LDLs to be low, our LDL cholesterols to be low. And we've seen that in some cases, that's the case. So what you're seeing in the bottom part of this figure is that we had sort of historical data for some of these people. We haven't done a lot of lipid analysis in the trial. So in some cases, the LDL level had been measured pretty far back in the past. We didn't have any enrollment week data points. We didn't have data points two and three. but we can compare at least the legacy values of HDL to, I mean, to LDL to what we had very quickly in the trial and saw that the levels had come down significantly.

So we're seeing all kinds of things like this. You know, I don't have time to go through all of the data, but this is another encouraging sign that we may be improving cardiovascular health at least in a subset of our population.

So we discovered some new things also that seem like they likely to have some generality to them. Again, these are just three examples. And this happens to be in men, but we've seen trends like this in women as well. So blood urea nitrogen. So I've already showed you that kidney function seems to be improving in TRIIM-X just as it did in TRIIM measured by the eGFR. But we're finding this new effect independent of eGFR. So even people whose eGFRs have been fairly stable and not improving sometimes show a reduction in their blood urea nitrogen level.

And I'm very intrigued by this because my interpretation of this is that this is a sign that you're taking nitrogen out of the blood and putting it into tissues. You're building up your lean body mass. Blood urea nitrogen is a consequence of amino acid turnover and breakdown. It's a waste product of amino acid metabolism. And to pull it out of the bloodstream independently of urinating it out implies that it's going into new proteins, making new proteins in the body.

So very early stuff here, but I think kind of provocative. And we'll see how this goes over time.

Here's another one that I was very interested in, lung function. And so I personally seem to have some kind of issue with gas exchange in my lungs. I've been looked at. And so I'm kind of conscious of CO2 levels. If your CO2 levels go up to around 30 or 33, you're beginning to get above the normal range, which means that you're starting to have impaired lung function. And this tends to get worse and worse as we get older and older.

And so here's an example in four different cases two of them females and and two of the males in which the CO2 levels seem to be going down pretty quickly with respect to the onset of the treatment and yes the data bounced around a little bit but you can see that the trends are pretty consistent so it possible that in addition to the other things that we observed in TRIIM that We actually observing here an improvement of pulmonary function improved lung uh function gas exchange function that would be a very exciting new development to add in to the the list of benefits of the TRIIM treatment and all is very very new so um

Persistence of TRIIM Effects and New PhenoAge Data

we also had the opportunity to check on something else that's been um you know in the back of my mind for a long time, and that is how long do the epigenetic aging reversal effects of TRIIM last?

So we saw in TRIIM that six months after the end of treatment, in some of the epigenetic aging clocks, there's a little bit of backsliding back toward sort of catching up with your chronological age. But in GrimAge, for example, there was no backsliding. Six months after the trial, you were just as young relative to your chronological age as you were at the end of the treatment.

But we had an opportunity here because one of, we actually have two of the guys who are in TRIIM are now in TRIIM-X. And we have this data for one of them. I was able to sort of reconstruct what happened to him over the last six years.

So he finished his treatment six years ago. Steve Horvath went back and back calculated his PhenoAge plasma level, which we're calling PhenoBioAge on the right, or PBA. So pardon me for the inconsistency and nomenclature here, but we're talking about the same thing.

So looking at this particular volunteer, based on the PhenoBioAge or Pheno plasma age, at the beginning of TRIIM, He was 53.6 years of age, but his PhenoBioAge was 55.2. So based on that clock, he was actually 1.6 years older biologically than he was chronologically at the beginning of that trial.

Now, if you look at Steve's calculations on the left, you see that he went from that starting age of 55.2 based on PhenoBioAge down to a biological age of about 42 at the end, which is a gain in epigenetic age of 13.2 years. So you know that in TRIIM, people reversed by on the average about two and a half years, but some people did better. And this is an example of that sort of thing.

So, because he's now the same guy, and he's now in the new trial, and we can still calculate his PhenoBioAge in the same way that we did before, we can now look and see where he started TRIIM-X and he started TRIIM-X with a PhenoBioAge of 2.6 years younger than his chronological age. So if you take into account the fact that he should have been at least 1.6 years older than his chronological age based on what we saw in TRIIM, he's still 4.2 years better off now than he would have been if he had not been in the trial at all. And this is six years after stopping treatment.

So this is encouraging. This implies that this may actually last for a while. Of course, it's only PhenoBioAge. It's not all of the epigenetic aging clock. It's not DNA methylation. And so it's preliminary data, but it indicates that maybe not only can we reproduce an anti-aging effect in TRIIM-X because we're actually seeing... Well, actually, I'm jumping the gun a little bit. You'll hear about that in the next slide. Suffice it to say, we can still detect the effect that we induced six years earlier in this one volunteer. That's encouraging.

So here's the second aging clock result that we have to show out of the TRIIM-X. And again, this is all based on Pheno plasma age. So what you're seeing plotted on the y direction is the difference between Pheno plasma age and chronological age, and same data representations you've been seeing.

So before treatment, you know, to the left of the vertical dash line, you see pretty stable or even increasing differences, which vary quite a bit from volunteer to volunteer, but as in TRIIM, our people were pretty healthy, and most of them have biological ages, at least based on this measure that are younger than their chronological ages.

But as we begin treatment we seeing in these three cases at least that the Pheno plasma age is beginning to regress almost immediately. And this is interesting because this is spread out over both males and females and over age ranges. So the older female is up at the top. She's near the upper limit of age for people who are eligible to enter the TRIIM-X trial. she was going in the wrong direction, if anything, at the beginning, but then she just showed this nice, steady improvement over time in her Pheno plasma age afterwards.

And then compare her to the younger female who started off ahead of the game by about 11 years or so. But in spite of that great advantage going into the trial, she rapidly went down to about minus 15 from minus 11. And so she picked up four years of Pheno plasma age in a very short amount of time.

Now, what we're seeing more generally is that people don't respond this fast usually, but these people are responding pretty quickly. I think that we're going to see a different pattern in other people, which is going to be very interesting in and of itself. But the other data we have is consistent with this data showing that it looks like even in TRIIM-X, at least if you go on the basis of this one sort of do it on the fly kind of clock, we're able to see evidence of aging reversal once again.

Conclusion of Presentation

So that's the preliminary results that we have so far. So far, everything that we can measure seems to be going in the right direction. So we're encouraged.

So in conclusion, you know, things are going pretty well. And it looks like we're reproducing the TRIIM trial results. But of course, it's going to take years before we can really prove it. This is all sort of very preliminary stuff, but it's I'd rather have preliminary results that look like this than that don't. So I'm really happy to share this with you and to share with you the news. also that it looks like even in ways that we didn't know before that we didn't see and TRIIM itself maybe because we just didn't think about it at the time that aging seems to be going in reverse in these new ways as well so um i i hope that that was illuminating to some of you and uh i sorry if i went to uh too much of the background but I wanted to bring everybody up to speed So thanks a lot for your attention And if there any further questions, if there's any other questions that you have, I'd be happy to discuss it.

Thank you so much, Greg. You have no idea how many questions you have. They were piling up. We have a few uploaded and downloaded. And maybe a few people have to drop on anyways because we're at the hour but i want to check would you be able to stay on a few more minutes or sure sure yeah okay well let's see how many we get through let us know in case you have to drop off and just be very outspoken with that we won't we won't even okay should i stop i will get started with the first question i may stop sharing your slide just for now so we can all see each other but we may get back to it uh eventually okay but thank you this was incredible

Q&A: Controlling for Confounders (Andy)

First question without any ado from me. Andy.

Yeah, great talk, Greg. I was curious with the cohort being biohackers, have you controlled it all for other things like taking NR, rapamycin or fasting? Are you tracking those to see if you get additive effects?

Yeah, we, those are ongoing issues with a lot of people. We have some people taking mass quantities of NR or NMN. We have people taking a lot of resveratrol. um actually you guys may know this but the evidence in favor of resveratrol has fallen apart recently and uh so uh to some people who are fairly enthusiastic about resveratrol i've been sending out this video uh that just demolishing the idea that it actually helps uh with uh with uh you know sirtuin function in humans um

so yeah and then there's this constant tension between what we're trying to do and CR. A lot of the people in our trial are pretty skinny and they are really CR devotees. There's one CR guru who signed up for this thing at the beginning, but then when I told her she has to sort of back off on CR, she decided not to. She just couldn't part with it. But other people are working with me on that because CR is a catabolic state, right? But by depriving yourself of energy, you start to tear down your own cellular protein stores and energy stores to give yourself the energy that you need to stay alive And that good because it tears away old damaged molecules But what we want to do is the opposite We need to grow you We need to build you up. We need to increase your lean body mass. We need to give you new cells in your thymus. And that's all inhibited by calorie restriction. Calorie restriction in humans is immunosuppressive. And so we can't really have too much of that.

On the other hand, we don't want to deprive people of the benefits of it. So we're trying to find a way to sort of optimize the balance between those things. And I can't say that we've exactly found that yet, but we're working on it.

Wouldn't that suggest a cycling? I mean, a cycling of autophagy and refeeding and a cycling of growth hormone and calorie restriction mimetics rather than simultaneously doing all of it?

Yeah, you really don't want to do it simultaneously. So I point out to the guys and gals in the trial that, you know, calorie restriction is good, but thymuses are good too. So, you know, you don't have to do one thing all the time. You know, you can go from one thing to the other. It's like the song, you know, to turn, turn, turn. There's this to everything. There's a season. You know, there's a time to build up. There's a time to break down. Right. there's a time to be on growth hormone, there's a time to be on CR.

So I think that you have to use your time in TRIIM-X or, you know, if you're doing TRIIM-X to maximize your TRIIM-X benefits. And if you take a holiday from CR for even a year, you're not going to age by more than a year anyway. But the fact is you're actually going in reverse. So the thing about the TRIIM protocol is it actually makes aging go in reverse. Calorie restriction does not make aging go in reverse. calorie restriction slows aging down. It does not reverse it. So you can be on calorie restriction all day long and you will live longer, but you will not stay young indefinitely.

So what we want to do is to make sure that you get the benefits of TRIIM-X. And then to the extent that you can combine that with a little bit of CR, it's a good thing. And we want to allow that to happen, but we're still working out the correct balance.

All right. Well, we need to make such a song.

Q&A: Protocol Optimization

Okay, Steve, you're next.

Yeah, great presentation. My question is about your speculations as to altering the protocol to make the growth hormone administration less frequent, possibly to remove some degree of growth hormone resistance that might be induced, but also to make it, let's say, medically or commercially more acceptable for people. And in terms of, you know, the combo therapy, do you think that your protocol of administering them all at the same time would be the way to do it? Or do you think that there may be some different kind of protocol where the growth hormone and the metformin and the DHEA might be administered on different protocols?

I'm not totally sure I understand, but yeah, so the real question is, do you give things like metformin and DHEA when you're not giving growth hormone? And I don't see any good reason not to, for the most part. There are some hazards of both. Sometimes you can go wrong with metformin, Sometimes you can go wrong with DHEA. So you have to be a little bit smart about it. But those hazards are probably no different whether you're taking growth hormone or not taking growth hormone.

So in TRIIM, we ask everybody to take all these things simultaneously four days a week and not to take them on the other days just to keep things simple and to and to reduce cost. but the cost of these things is really minimal so that the cost issue is very minimal.

And so in TRIIM-X, some people are taking these things more frequently than four times a week and some people are not. And we're beginning to reconstruct some information about which way most people should go and exactly how you adjudicate that. But we're still in the process figuring all of that out.

But your point is well taken that we're using metformin and DHEA for a specific reason in our protocol, and that's to block the diabetogenic effects of growth hormone, but that makes perfect sense to take them at other times as well because they have independent benefits. So we're going to have a lot more information about that. We actually have some preliminary data which I find very interesting which I not going to go into more detail about right now But yeah there something to be learned there It definitely something to be learned there That a great question Great question Steve yes Thanks.

Q&A: Economics and Investment

Tom?

Yeah, this is an economics question, which is whether or not investors are willing to support more expensive, larger clinical trials for drugs that are off patent. And there's a number of economists like Susan Athey at Stanford, who have written both about this problem, but also different approaches to solving that market failure.

So you'd like me to just comment on that? Yes.

All right. So here's the ultimate comment on that. Actually, there's two major ways of addressing that issue. One is to create new pharmaceuticals so they're not off patent. and two is to create work around so you don't need the high cost items so that the economics of it work out much more easily.

We're actually working on both of those. We've worked a fair amount on finding alternatives to growth hormone, which is the expensive item in our cocktail. So far, I'm not satisfied with those results. So we're not really ready to go down that pathway yet, but we're continuing to investigate that. We're probably going to be getting more information on that with some volunteers later this year.

But there's another way, and that is to make the growth hormone ourselves. So, you know, growth hormone may be off patent, but every time you make a new variation, you have to go through another approval process with the FDA. And growth hormone is not perfect. So there are certain features of growth hormone that could be improved without actually changing the molecule itself.

If you change the stabilizing solution that the growth hormone is in, you may be able to gain some market advantages, some matters of convenience for the consumer that would allow a new patent and that would allow that proprietary aspect to be overcome.

And it turns out that I'm a cryobiologist in my day job. I preserve kidneys at cryogenic temperatures and things like that. And so I know something about how to stabilize proteins And I got some ideas that nobody had before And I think that we can come up with a version of growth hormone that we can make ourselves about five times less costly than what is commercially available now that will be superior to commercially available growth hormone

And we have some people that are interested in helping us do some pilot studies to prove out that concept. If we can do that, then we'll own that growth hormone version. We will not have a patent problem. We will have a massive selective advantage over any potential competitor in the sense that nobody will be able to offer the same price that we can offer to our own treatment clients.

And so I think that that could unlock the door to tens of millions of people getting our treatment, because then it would be much, much, much more affordable, and all of the marketing issues will be overcome.

So that's a major thrust of our interest going forward. Unfortunately, we have very little money in the bank right now, but we have enough that we should be able to move forward on that fairly soon. And we have a couple of people we're talking to about that.

So that's, I think, what the ultimate solution is. And that's the future of our company, I think. And then if we can develop lower cost alternatives, we will do that too. There's one wrinkle there, one particularly interesting possibility there that no one else has been pursuing that we're going to hopefully get some data on in the next year or so. And if that happens to work, it'll be really, really interesting. And that's all I can tell you about it.

So are you actively looking for investments at the moment? Are you actively taking on investors or funders?

Yeah, we're always looking for investors, but unfortunately, the investors are not always looking for us. So, and it's for just the reason that Tom brought up, you know, they say, well, you know, growth hormone is out there, you don't own it, you know, et cetera, et cetera. And we have a patent application on our treatment, but people worry about bootlegging and things like that.

So we're growing our company by organic growth, even though this is a trial, we're allowed to make a small amount of margin up of each person in the trial beyond the cost. So we have our administrative needs covered. And we're actually demonstrating that there's a market for this product, even despite the high cost. And nobody's sort of going in the competition against this so far. At least not that we been told

And so I think that we have a way forward through organic growth And that will eventually lead to these new products that I was just discussing But if somebody wants to jump in at the strategic moment in time, I think it could be a super good investment. That was my opinion.

All right. Duly noted.

Q&A: Soft Tissue Growth

Okay, Raphael, next question.

Hello, Greg. Thank you for the presentation. I have a question. We know that there are some soft tissue like nose and hair that continue to grow throughout life. And I was wondering if you think that growth hormone has an effect on this as it would increase this growth or not at all?

The growth of hair? The growth of? You're saying the growth of hair? oh no uh here uh the nose for example the nose or hairs i think they are going you know and do you think this could be increased by uh GH from on injections

so let me just make sure that i understand the question i'll restate it and then if i restate it wrong you can correct me okay rafael so uh rafael is saying that there there are things in in the body that go on growing throughout life. And one reason that we look older is not necessarily because we're falling apart, but it's because things keep growing. So older people have big ears and they have big noses and they tend to have big chins. And it's because your nose never stops growing and your ears never stop growing. And so if you take growth hormone, are you going to actually look older because you're looking more like, you know, a big nose, big eared old person.

And I think the time will tell on that, but we haven't really noticed anything like that. I will say one thing, and that is that even though all of the evidence says that we're reversing aging, people are not really looking younger facially so far. And we may change that.

I don't know if you guys have seen these advertisements on television for this product called CeroVital. Kim Douglas comes on and says, oh, I heard about this youth hormone and it even gets rid of wrinkles. But you know, we're just not seeing that. And actually, I think their product doesn't work.

So I think that we're not quite at that point yet. But we may get there if we continue to refine things. We actually we're not quite at that point yet. But we may get there if we continue to refine things. We actually have some interesting ideas in mind that may be helping us out in that regard.

But all I can say is that we didn't notice anybody's noses or ears or nose hairs or ear hairs or things like that getting worse in TRIIM. And so far, nobody's complained. Actually, there's one person in TRIIM-X who I will not refer to by name here, who is suffering from the opposite problem from excessive hair growth. There's actually a problem with hair loss, but I don't think that's necessarily related to the TRIIM-X treatment. I think that's an ongoing problem.

So I don't know if that answered your question, but that's about as good as I can do on it, I think.

I just wanted to quickly jump in on that, Greg, follow-up question. On that subject, are you gathering daily facial pictures? Because I'm usually interested in that for measuring aging, but yeah based on what Raphael just said it's like a double reason to want to collect yeah yeah yeah so uh we have taken photographs of people at baseline and we took a lot of pictures of people's hair at baseline also uh because we don't want to miss out on a hair darkening effect this time like we like we did last time uh so yeah that is part of the piece yeah thanks okay no problem next one up with Gail

Q&A: Kidney Transplants and Thymic Reprogramming

Gail, hey, how are you? Hi, I'm not now. Great talk. Yeah, yeah. So exciting that you're into phase two here. I have a question that has a finance market implication, and that question is this. So, You mentioned improvement in GFR and glomerular renal function. And the question is, would this protocol be compatible with somebody who's had a kidney transplant and is therefore on immunosuppressants? Because if you could extend transplant life by a year, okay, by two years, you're going to be generating a short-term immediate, besides for the health benefit, financial benefit that's going to tie right into Medicare You got a potential financing opportunity here So what the deal What do you think?

That's a really smart question. So I'd like to sort of amplify that a little bit. So there's all kinds of ways in which these subjects may intersect, kidney transplant subject and what we're doing.

one thing is consider the cost of the TRIIM-X uh treatment and all the monitoring it's pretty expensive for a lot of people it's 18 000 a year but consider the cost of a kidney transplant yeah you're talking you know more like half a million or something like that and then consider the cost of dialysis every year which is you know in excess of 18 000 for a lousy quality of life in which you're dead about five years later on average uh so yeah, if we could marry these things, the cost aspects of the TRIIM-X treatment, even with all the monitoring thrown in, which is a big part of the overall cost, becomes negligible in the context of treating somebody with a kidney transplant.

So that's one aspect of it. Another aspect of it is that growth hormone seems to benefit kidney function. So if you have a transplant is not quite up to speed, but we might be able to even compensate for that deficiency and help you in that way as well.

Now, there's two then competing additional considerations that have to be factored in. One is that, as you know, the reason that we're giving the growth hormone in the beginning is to regenerate the thymus, and that's to improve immune system function. As you pointed out, if you have a kidney transplant, you're on immunosuppression. And so there may be a bit of a tension between those two things. So that's the first aspect is that by improving immune system function, you may accelerate transplant rejection, and that would be a problem.

But the other consideration is this other lofty goal, which I've had in mind for the last 20 years or so, which we haven't had a chance to work on yet. But again, this is another thing that investors can help us with a lot. And that is that there's a way, and Gail, you've heard this story because Gail was sitting in the room when I first introduced this concept of the TRIIM treatment to the world at a meeting in the Bay Area called the Health Extension Salon I believe

And I mentioned at the end of the talk that there something that I called thymic magic at the end of the talk. And the thymic magic aspect has to do with the fact that the thymus actually has two jobs. It manufactures cells that destroy enemies, and it also manufactures cells that do not destroy friend, in other words, self.

And it's that second aspect that's just really interesting because if you have a thymus, which most of us don't, you know, to speak of after the age of 30 or so, but if you have a decent amount of thymus, you can actually re-engineer the thymus to accept any graft as self and not reject it. And this has been shown in every animal model there is of transplant and it always works, including in large animals. And it seems like something that would be doable in humans based on what we saw in TRIIM. In other words, in TRIIM, we saw we were able to regenerate the thymus. That means that there's some place to do this engineering work on.

It involves a minor surgical procedure to introduce the right antigens into the thymus. But once you do that, just give it a few months and your transplant rejection should be eliminated for life. You can then go off immunosuppression for the rest of your life and keep that kidney as though it were from your identical twin, which means it will last for 25 years instead of for five or 10 years.

but it actually gets better than that because you can then apply the same technology to curing all autoimmune diseases for exactly the same reason because an autoimmune disease is when the body forgets that you are you and it starts attacking you but you can reprogram the thymus to recognize that that thing that is being attacked is actually you and delete all of the cells that engaging in the attack.

This also has been proven to work in every animal model has been tried in, which are a large number of them. And nobody has tried this in humans, just as nobody has tried the transplant rejection in humans yet either.

So if we succeed as a company these are other multi dollar areas that we want to get into And again that would be a nice place for investors to jump in here I actually run this company called 21st Century Medicine during the day. We have a pig colony in the building, so we can do pig experiments. And there's a lot of interesting immunological work done on pigs. So we could actually test this rejection issue in the pig model with relatively little overhead because the colony already exists and I run the lab so I don't have to charge my other company that much money to do these studies.

So, you know, that's another potentially highly strategic way that actually having investment in the company could help us, which will take us another 20 years to raise the money at the rate we're going at right now. But of course, I expect that to accelerate. But just for any investor types out there, these are not fantasies either. I mean, the animal studies are unequivocal. They're numerous, both with respect to transplant rejection and autoimmunity.

It's just that nobody has bothered to try this in people for reasons that escape me, just as nobody bothered to try to regenerate the thymus in humans until we came along, except for a few brave doctors working on HIV patients because their patients were dying in front of their faces. And even they have seemingly given up on that process because they're worried about their patients getting cancer, which they haven't gotten cancer, but even that has been sort of dying out.

So somebody's got to do these things and darn it, we're going to do it if nobody else does.

We have an investment number for those individual procedures. Just a rough ballpark. We won't hold you to it.

Well, we're looking for at least a million, but of course that's nothing. So, you know, we could use five or 10, but you know, that all depends on the investor and what they're interested in and so on and so forth and investment terms, which we think we can make favorable. So we would, we, we would talk to anybody who's interested, but just put it that way.

Very nice.

Q&A: Bias and Economics

Okay. This is the information that I wanted. We have Matt with one more question, and then I want to have one more wrap up questions of how to perhaps how this group could help you moving forward. But Alan, questions next.

ALAN LEVINEY- Here there, can you hear me OK? ALAN LEVINEY- Yes. ALAN LEVINEY- Yep. ALAN LEVINEY- Great. So two questions. Thank you. Thank you. Alex, questions next.

Hey there. Can you hear me OK? Yes. Yep. Great. So two questions. Hopefully I can sneak that in and Allison's OK with that.

The first is that we all want this to work. And I want to make sure that we're not engaging in unconscious cherry picking or motivated result seeking. And so I'm curious how you, as you've approached this, have made sure that you're not like falling into your own unconscious bias that you want it to work.

Yeah, it's always a problem. It's always a question. But in the end of the day, you analyze the data and the data are what the data are. And there's not a lot you can do about it. So I'm I'm aware of that, you know, potential bias. I can't swear that, you know, could not affect me, but I'm not aware of it being an issue right now. So that's about all I can say.

I guess there's one thing that I should bring up here because I never get a chance to mention this. And it kind of is up that tree, you might say. So I've been very impressed with the lymphocyte to monocyte ratio result that we got in TRIIM. and partly because of the story that it tells about the NAD connection between what we're doing and aging.

So if we increase the lymphocyte to monocyte ratio by reducing CD38 positive monocytes, then we should increase tissue stores of NAD and we should rejuvenate the body and get the results that we are seeing.

And by the way, as far as the epigenetic aging reversal is concerned, Steve Horvath did all of those tests for us. I didn't touch any of that. And the Pheno plasma clock results that I showed you, it's all based on mathematical formulas that cannot be fudged and that I do not do myself. So there's no bias in any of that. uh that steve uh horvath is about the most non-biased person you'll ever run into yeah thank you just to tell everyone i i will do the epigenetic clock analysis i'm incorruptible whatever comes out comes out you know that true that is true and i i kind of blinded you know so yeah yeah he very objective Yeah that one thing we like about him He has no susceptibility to bias. I think he just lets the chips fall where they may.

But to complete the story, you know, so I may be a little bit biased in this respect, so that I want this lymphocyte to monocyte ratio to mean something, right? Because the story makes so much sense, right? And we had highly statistically significant results. There's no doubt about it.

But there's this bizarre aspect of this thing, which I've never had really a chance to explain to anyone. So here's true confession time. You do not see an increase in lymphocyte to monocyte ratio if you measure it with ordinary techniques. You only see it if you measure it with these ultra sophisticated techniques that are used at Stanford, the CyTOF method.

And I'm still hitting myself in the head trying to figure out how it's possible for there to be a discrepancy between these things. But I haven't quite figured it out yet. So maybe I'm doing something wrong and analyzing the conventional blood data. So I continue to present that result because I don't think it can be wrong because the statistics don't lie. And then on top of that, we have the connection to NAD, but it could be wrong in some bizarre way. I don't know how. I really cannot explain how it could possibly be wrong.

But if you have a discrepancy like that, you need to check your premises. And so that could be an example in which I want that to be true. So I've been presenting that without really being good enough about presenting that caveat. So now I've come clean on that.

But as far as everything else is concerned, I think I'm pretty much objective. And then I did show you results, preliminary results for TRIIM-X that were favorable. I didn't show you any results that were unfavorable. There have been some results that are not as favorable, but we're so early into the trial. I don't think they mean anything yet.

And the results that I did show you are supported by other results that are less obvious. So I think that It's right. But again, as I told everybody, we're less than six months into this trial. We've got another year and a half to go, probably, before we have all of the data in. Very limited number of people blah blah blah We only have one fourth of the population that we shooting for one third to one fourth So it all very early stuff right I just wanted to give you a flavor of what we seeing what the trends look like things that are exciting to me and hopefully will all pan out. But I'm not seeing any red lights. I'm seeing things that look favorable globally. So I think that much is objective.

Nice.

So I'm curious, if results are neutral to positive, is there a structure that allows the trials to be self-funding, where essentially anyone who wants to join the trial can pay for all infrastructure costs and you deliver it at cost? Or is that something that the FDA thinks creates a moral hazard and therefore it's not allowable?

No, there are provisions at the FDA for that kind of trial. As a matter of fact, TRIIM-X is that kind of trial because we do not have investment money this time. So everybody in our trial is paying their own way. And one thing that that leads to is very committed volunteers. So you get very good behavior and you get very good communication from the volunteers. They want it to work out and they're extremely interested in the results. So that's all good.

And like I say, we're able to make a little bit of margin on each person. So our bank reserves are increasing slowly, not a lot, but enough to keep us alive as a company and allow us to pay the bills. And that's all allowed by the FDA. And as a matter of fact, the trial plan that we submitted to the FDA is on this basis and they approved it. So So TRIIM-X is going forward under our original IND with the FDA. And it is also under approval of an IRB that is aware of that cost sharing aspect of the study. So, yes, so we can expand that.

One beauty of this mechanism of funding is that since it's self-paid, there's no limit on the number of people we can enroll in the study. We can make the study as large as, you know, as there is a demand for people to enter it up to some point. You know, at some point, it's going to have to be converted from its trial into something else. But we're not close to that point at this point, I think.

So if the FDA knocks on our door you know a few years from now and says hey you know you got 100 people that have had this treatment They all paying you We think you know this is no longer a trial you need to ask for approval Well then we ask for approval at that point but we have data for 100 people So we should easily get the approval at that point. So there's a limit to it. But for the time being, for the near future, this mechanism functions for us.

Sounds great. Yeah. By the way, that's one that's just a very, very quickly. I know time is going by, but that's another area for investors to participate because we can convince people to pay for getting actually a treatment, but it's hard to pay for people to, it's hard to get people to agree to pay for being controls and not getting the treatment.

And so if we had some investment money coming in to cover the cost of controls, so we could run more controls. So we're looking at different kinds of control endpoints to try to find a way that makes it attractive for people to be controls. But if we had a little bit of investment to cover their costs, that would be even better.

Closing Call to Action

All right. Okay, so I just want to make sure we finish with, without letting you go, without you telling us what this group of people listening could help. And we've already heard a little bit about investment, But let's say I'm guiding other people to this talk and I'm saying just if you look at one thing, then look at the last minute. Then, you know, I guess what would you say in terms of do you need specific participants? Do you need specific types of investment? If there was like, you know, one wrap up, like a call to action that could help. I would really love if we could have that in a concise way at the end, which would be now.

Thank you, Alison. I really appreciate that. And that is a that's a unique feature of these seminars that you put on. nobody does this you always do this you always ask the speakers how you can help you know that's fantastic and highly appreciated and so uh i have you know two simple answers and basically the one the one answer is tell your friends and and let them know that TRIIM-X is open uh for enrollment because the way that we keep things moving forward is by enrolling more people in trial. It's our only source of income right now, but it's also our only source of knowledge.

So anyone who is interested in the trial and can pony up this $18,000 price tag, which is pretty steep. I agree. I know we're working on it, but anyone who's able to get to that sort of thing will be helping us and will be appreciated.

And then the other thing would be tell not just your friends, but tell your investment friends also, just in case they may want to take a shine to one of these other things that we talked about, either the growth hormone or the immunoengineering aspects or anything else that they may be interested in.

And best way to contact you would be to go on Intervene Immune's website?

Yes. So my, yeah, https://www.interveneimmune.com/ it's hard to forget. And fahy@interveneimmune.com. Also pretty hard to forget as long as you know my name.

All right. Well, Greg, I cannot thank you enough. This was really, really fantastic. I can't tell you how excited people were to have you on. I'm hoping that we hear from you again maybe in a year or so for another update with much more exciting data. Yes. Much more funding and having brought the price down then to negligible amounts. So maybe all went through it at this point. So thank you so, so, so much. This was really, really, really valuable for all of us.

Intuitions

• Thymic centrality to aging: Thymic involution drives T-cell depletion and immune decline, accelerating mortality; regeneration reverses this and non-immune aging hallmarks (e.g., hepatic tetraploidy, beta-adrenergic signaling, insulin sensitivity via thymus transplants in animals).

• hGH-DHEA-metformin synergy: hGH anabolically regenerates thymus/lean mass across species but induces insulin resistance; DHEA physiologically counters this (mimicking youth), metformin provides broad sensitization; intermittent dosing (4x/week) optimizes while minimizing costs/resistance.

• Multi-pathway aging reversal: Independent effects—thymic regeneration boosts naive T-cells/RTIs, reduces PD-1; monocyte depletion elevates tissue NAD+ (reversing mitochondrial aging); clocks show accelerating reversal (4x faster late-trial), persisting 6+ years.

• PhenoAge plasma clock trick: Real-time, blood-based proxy (no DNAm needed) replicates full epigenetic reversal (2.5 years avg), enabling intra-trial monitoring.

• Self-funding scalability: Participant-paid trials (IND/IRB-approved) generate data/margin organically, bypassing investor hurdles for off-patent drugs; targets n=80+ for robust stats.

• Thymic reprogramming: Regenerated thymus enables antigen-specific tolerance induction, potentially curing transplant rejection/autoimmunity (proven in diverse animal models, untried in humans).

• Proprietary hGH: Stabilize via novel cryopreservation-derived formulations for 5x cost reduction/new patents, enabling mass access.

• Cycling confounder management: Alternate anabolic (TRIIM-X) with catabolic (CR/autophagy) phases to avoid antagonism.

Transcription errors?

• "CRIM trial" → Corrected to "TRIIM trial" based on context (Thymus Regeneration, Immunorestoration, and Insulin Mitigation).
• "TRIMx/TRIM-X/TRIIM-X" → Standardized to "TRIIM-X" (extension trial).
• "pheno age/PhenoAge/Pheno plasma age/PhenoBioAge/PBA" → "PhenoAge" (standard clock); "PhenoAge plasma clock" or "Pheno plasma age" for blood-based component.
• "recent thymic immigrants" → "recent thymic emigrants (RTIs)".
• "Cytoff/CyTOF" → "CyTOF" (mass cytometry).
• "Keith Kelly" → Likely "Keith Kelley" (1986 rat study author).
• "Nir Barzilai" → "Nir Barzilai".
• "Steve Horbath/Steve Horvath" → "Steve Horvath".
• "Irina Convoy" → "Irina Conboy".
• "Bobby Brooke" → Likely "Bobby Brooks" (CEO; minor uncertainty).
• Lymphocyte:monocyte discrepancy: Self-noted by speaker (CyTOF vs. standard); potential artifact unresolved.
• Minor audio garbles (e.g., "juggling items", "uniformatized"): Interpreted as "optimized doses", "standardized".
• No major chemical misspellings; terms like IGF-1, NAD+, PD-1, PSA, eGFR, CRP standardized. Company: Intervene Immune (interveneimmune.com).

See also

• longevity
• ?thymus